An elevated number of differentiated osteoblast colonies can be obtained from rat bone marrow stromal cells using a gradient isolation procedure

G. Lisignoli, G. Remiddi, L. Cattini, B. Cocchini, N. Zini, M. Fini, F. Grassi, A. Piacentini, A. Facchini

Research output: Contribution to journalArticle

Abstract

Bone marrow stromal cells (BMSCs) for osteoblast differentiation studies can be obtained by gradient isolation techniques or by directly plating a filtered cell suspension. We compared these two procedures to evaluate whether this step is critical in order to obtain a high number of differentiated colonies. Isolated primary rat BMSCs were cultured in vitro with or without insulin-like growth factor II (IGFII), basic fibroblast growth factor (b-FGF), epidermal growth factor (EGF) or transforming growth factor β1 (TGFβ1), and histochemically and biochemically analysed at different time points. The gradient procedure produced a significantly higher number of colonies capable of osteoblastic differentiation. The growth factors had different effects. In particular, b-FGF and EGF significantly increased the number of Alizarin red S positive colonies, while IGFII and TGFβI exerted inhibitory effects. Nodules obtained on day 21 showed some alkaline phosphatase positive cells and were Von Kossa-positive. These data demonstrate that more differentiated colonies are obtainable from BMSCs isolated by the gradient procedure.

Original languageEnglish
Pages (from-to)49-58
Number of pages10
JournalConnective Tissue Research
Volume42
Issue number1
Publication statusPublished - 2001

Keywords

  • Bone marrow stromal cells
  • Differentiation
  • Growth factors
  • Isolation procedure
  • Mineralization

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Nephrology
  • Cell Biology
  • Orthopedics and Sports Medicine

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